中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2009年
6期
452-457
,共6页
贾洁爽%梅长林%付莉莉%戴兵%胡惠民
賈潔爽%梅長林%付莉莉%戴兵%鬍惠民
가길상%매장림%부리리%대병%호혜민
p38丝裂原活化蛋白激酶类%多囊肾,常染色体显性%表皮生长因子%罗格列酮
p38絲裂原活化蛋白激酶類%多囊腎,常染色體顯性%錶皮生長因子%囉格列酮
p38사렬원활화단백격매류%다낭신,상염색체현성%표피생장인자%라격렬동
P38 mitogen-activated protein kinases%Polycystic kidney,autosomal dominant%Epidermal growth factor%Rosiglitazone
目的 探讨罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶(MAPK)信号通路的作用.方法 分别用罗格列酮(RGZ,10 μmol/L)、过氧化物酶体增殖物活化受体γ(PPARγ)抑制剂GW9662(10 μmol/L)、RGZ(10 μmol/L)+GW9662(10 μmol/L)、p38MAPK特异性抑制剂SB203580(10 μmol/L)、SB203580(10 μmol/L)+RGZ(10 μmol/L)处理体外培养的多囊肾囊肿衬里上皮细胞(PKD细胞)2 h后,再用表皮生长因子(EGF)刺激不同时间,另设置空白对照组和单独EGF刺激组.采用Western印迹方法检测p38MAPK、磷酸化p38MAPK(p-p38)、增殖细胞核抗原(PCNA)表达;RT-PCR检测p38 mRNA表达;免疫细胞化学检测c-fos和c-jun的表达.结果 (1)与空白对照组相比,EGF显著上调p-p38、PCNA、c-fos和c-jun的表达(P<0.01).(2)与EGF单独刺激相比,RGZ显著降低p38活化和基因表达(均P<0.01).RGZ组、SB203580+RGZ组p-p38、PCNA、c-fos、c-jun表达明显下调(P<0.01),两组间差异无统计学意义.(3)与RGZ组相比,RGZ+GW9662组部分阻断RGZ的下调作用(P<0.05).结论 罗格列酮抑制多囊肾囊肿衬里上皮细胞增殖的作用机制可能与其降低p38MAPK活性,继而抑制PCNA、c-fos及c-jun表达有关.这种抑制作用是部分非PPARγ依赖的.
目的 探討囉格列酮對多囊腎囊腫襯裏上皮細胞p38促分裂原活化蛋白激酶(MAPK)信號通路的作用.方法 分彆用囉格列酮(RGZ,10 μmol/L)、過氧化物酶體增殖物活化受體γ(PPARγ)抑製劑GW9662(10 μmol/L)、RGZ(10 μmol/L)+GW9662(10 μmol/L)、p38MAPK特異性抑製劑SB203580(10 μmol/L)、SB203580(10 μmol/L)+RGZ(10 μmol/L)處理體外培養的多囊腎囊腫襯裏上皮細胞(PKD細胞)2 h後,再用錶皮生長因子(EGF)刺激不同時間,另設置空白對照組和單獨EGF刺激組.採用Western印跡方法檢測p38MAPK、燐痠化p38MAPK(p-p38)、增殖細胞覈抗原(PCNA)錶達;RT-PCR檢測p38 mRNA錶達;免疫細胞化學檢測c-fos和c-jun的錶達.結果 (1)與空白對照組相比,EGF顯著上調p-p38、PCNA、c-fos和c-jun的錶達(P<0.01).(2)與EGF單獨刺激相比,RGZ顯著降低p38活化和基因錶達(均P<0.01).RGZ組、SB203580+RGZ組p-p38、PCNA、c-fos、c-jun錶達明顯下調(P<0.01),兩組間差異無統計學意義.(3)與RGZ組相比,RGZ+GW9662組部分阻斷RGZ的下調作用(P<0.05).結論 囉格列酮抑製多囊腎囊腫襯裏上皮細胞增殖的作用機製可能與其降低p38MAPK活性,繼而抑製PCNA、c-fos及c-jun錶達有關.這種抑製作用是部分非PPARγ依賴的.
목적 탐토라격렬동대다낭신낭종츤리상피세포p38촉분렬원활화단백격매(MAPK)신호통로적작용.방법 분별용라격렬동(RGZ,10 μmol/L)、과양화물매체증식물활화수체γ(PPARγ)억제제GW9662(10 μmol/L)、RGZ(10 μmol/L)+GW9662(10 μmol/L)、p38MAPK특이성억제제SB203580(10 μmol/L)、SB203580(10 μmol/L)+RGZ(10 μmol/L)처리체외배양적다낭신낭종츤리상피세포(PKD세포)2 h후,재용표피생장인자(EGF)자격불동시간,령설치공백대조조화단독EGF자격조.채용Western인적방법검측p38MAPK、린산화p38MAPK(p-p38)、증식세포핵항원(PCNA)표체;RT-PCR검측p38 mRNA표체;면역세포화학검측c-fos화c-jun적표체.결과 (1)여공백대조조상비,EGF현저상조p-p38、PCNA、c-fos화c-jun적표체(P<0.01).(2)여EGF단독자격상비,RGZ현저강저p38활화화기인표체(균P<0.01).RGZ조、SB203580+RGZ조p-p38、PCNA、c-fos、c-jun표체명현하조(P<0.01),량조간차이무통계학의의.(3)여RGZ조상비,RGZ+GW9662조부분조단RGZ적하조작용(P<0.05).결론 라격렬동억제다낭신낭종츤리상피세포증식적작용궤제가능여기강저p38MAPK활성,계이억제PCNA、c-fos급c-jun표체유관.저충억제작용시부분비PPARγ의뢰적.
Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt.
